Enhanced limonene production by metabolically engineered Yarrowia lipolytica from cheap carbon sources

ABSTRACT The genes encoding gluconeogenic enzymes in the nonconventional yeast Yarrowia lipolytica were found to be differentially regulated. The expression of Y. lipolytica FBP1 (YlFBP1) encoding the key enzyme fructose-1,6-bisphosphatase was not repressed by glucose in contrast with the situation in other yeasts; however, this sugar markedly repressed the expression of YlPCK1, encoding phosphoenolpyruvate carboxykinase, and YlICL1, encoding isocitrate lyase. We constructed Y. lipolytica strains with two different disrupted versions of YlFBP1 and found that they grew much slower than the wild type in gluconeogenic carbon sources but that growth was not abolished as happens in most microorganisms. We attribute this growth to the existence of an alternative phosphatase with a high Km (2.3 mM) for fructose-1,6-bisphosphate. The gene YlFBP1 restored fructose-1,6-bisphosphatase activity and growth in gluconeogenic carbon sources to a Saccharomyces cerevisiae fbp1 mutant, but the introduction of the FBP1 gene from S. cerevisiae in the Ylfbp1 mutant did not produce fructose-1,6-bisphosphatase activity or growth complementation. Subcellular fractionation revealed the presence of fructose-1,6-bisphosphatase both in the cytoplasm and in the nucleus.

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Biofilm formation by a biotechnologically important tropical marine yeast isolate, Yarrowia lipolytica NCIM 3589

Water Science & Technology ◽

10.2166/wst.2008.479 ◽

2008 ◽

Vol 58 (6) ◽

pp. 1221-1229 ◽

Cited By ~ 11

Author(s):

D. H. Dusane ◽

Y. V. Nancharaiah ◽

V. P. Venugopalan ◽

A. R. Kumar ◽

S. S. Zinjarde

Keyword(s):

Yarrowia Lipolytica ◽

Biofilm Formation ◽

Edible Oils ◽

Carbon Sources ◽

Three Dimensional ◽

Ecological Niches ◽

Lag Phase ◽

Biofilm Growth ◽

Water Media ◽

Biofilm Development

Biofilm formation by Yarrowia lipolytica, a biotechnologically important fungus in microtitre plates, on glass slide surfaces and in flow cell was investigated. In microtitre plates, there was a short lag phase of adhesion followed by a period of rapid biofilm growth. The fungus formed extensive biofilms on glass slides, whereas in flow-cells a multicellular, three-dimensional microcolony structure was observed. The isolate formed biofilms in seawater and in fresh water media at neutral pH when grown in microtitre plates. The carbon sources differentially affected formation of biofilms in microtitre plates. Lactic acid, erythritol, glycerol, glucose and edible oils supported the formation of biofilms, while alkanes resulted in sub-optimal biofilm development. A variation in the morphology of the fungus was observed with different carbon sources. The results point to the possible existence of highly structured biofilms in varied ecological niches from where the yeast is isolated.

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HOG-Independent Osmoprotection by Erythritol in Yeast Yarrowia lipolytica

Genes ◽

10.3390/genes11121424 ◽

2020 ◽

Vol 11 (12) ◽

pp. 1424

Author(s):

Dorota A. Rzechonek ◽

Mateusz Szczepańczyk ◽

Guokun Wang ◽

Irina Borodina ◽

Aleksandra M. Mirończuk

Keyword(s):

Transcription Factor ◽

Yarrowia Lipolytica ◽

Carbon Sources ◽

Hyperosmotic Stress ◽

Sensitive Strain ◽

The Other ◽

High Concentration ◽

Other Hand ◽

Yeast Yarrowia Lipolytica ◽

Better Than

Erythritol is a polyol produced by Yarrowia lipolytica under hyperosmotic stress. In this study, the osmo-sensitive strain Y. lipolytica yl-hog1Δ was subjected to stress, triggered by a high concentration of carbon sources. The strain thrived on 0.75 M erythritol medium, while the same concentrations of glucose and glycerol proved to be lethal. The addition of 0.1 M erythritol to the medium containing 0.75 M glucose or glycerol allowed the growth of yl-hog1Δ. Supplementation with other potential osmolytes such as mannitol or L-proline did not have a similar effect. To examine whether the osmoprotective effect might be related to erythritol accumulation, we deleted two genes involved in erythritol utilization, the transcription factor Euf1 and the enzyme erythritol dehydrogenase Eyd1. The strain eyd1Δ yl hog1Δ, which lacked the erythritol utilization enzyme, reacted to the erythritol supplementation significantly better than yl-hog1Δ. On the other hand, the strain euf1Δ yl-hog1Δ became insensitive to supplementation, and the addition of erythritol could no longer improve the growth of this strain in hyperosmotic conditions. This indicates that Euf1 regulates additional, still unknown genes involved in erythritol metabolism.

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Production of citric and isocitric acid by Yarrowia lipolytica strains grown on different carbon sources

Turkish Journal of Biochemistry ◽

10.5505/tjb.2014.92005 ◽

2014 ◽

Vol 39 (3) ◽

pp. 285-290 ◽

Cited By ~ 7

Author(s):

Fusun Bahriye Ucar ◽

Gonul Celik ◽

Onur Akpinar ◽

Cengiz Corbaci

Keyword(s):

Yarrowia Lipolytica ◽

Carbon Sources ◽

Isocitric Acid

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Metabolomic elucidation of the effects of media and carbon sources on fatty acid production by Yarrowia lipolytica

Journal of Biotechnology ◽

10.1016/j.jbiotec.2018.02.011 ◽

2018 ◽

Vol 272-273 ◽

pp. 7-13 ◽

Cited By ~ 5

Author(s):

Eun Ju Yun ◽

James Lee ◽

Do Hyoung Kim ◽

Jungyeon Kim ◽

Sooah Kim ◽

...

Keyword(s):

Fatty Acid ◽

Yarrowia Lipolytica ◽

Acid Production ◽

Carbon Sources ◽

Fatty Acid Production ◽

Effects Of Media

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The dimorphic yeast Yarrowia lipolytica possesses an atypical phosphofructokinase: characterization of the enzyme and its encoding gene

Microbiology ◽

10.1099/mic.0.27856-0 ◽

2005 ◽

Vol 151 (5) ◽

pp. 1465-1474 ◽

Cited By ~ 13

Author(s):

Carmen-Lisset Flores ◽

Oscar H. Martínez-Costa ◽

Valentina Sánchez ◽

Carlos Gancedo ◽

Juan J. Aragón

Keyword(s):

Yarrowia Lipolytica ◽

Carbon Sources ◽

Hill Coefficient ◽

Single Type ◽

Kinetic Characteristics ◽

Gene Encoding ◽

Dimorphic Yeast ◽

Encoding Gene ◽

Cooperative Kinetics

The phosphofructokinase from the non-conventional yeast Yarrowia lipolytica (YlPfk) was purified to homogeneity, and its encoding gene isolated. YlPfk is an octamer of 869 kDa composed of a single type of subunit, and shows atypical kinetic characteristics. It did not exhibit cooperative kinetics for fructose 6-phosphate (Hill coefficient, h 1·1; S 0·5 52 μM), it was inhibited moderately by MgATP (K i 3·5 mM), and it was strongly inhibited by phosphoenolpyruvate (K i 61 μM). Fructose 2,6-bisphosphate did not activate the enzyme, and AMP and ADP were also without effect. The gene YlPFK1 has no introns, and encodes a putative protein of 953 aa, with a molecular mass consistent with the subunit size found after purification. Disruption of the gene abolished growth in glucose and Pfk activity, while reintroduction of the gene restored both properties. This indicates that Y. lipolytica has only one gene encoding Pfk, and supports the finding that the enzyme consists of identical subunits. Glucose did not interfere with growth of the Ylpfk1 disruptant in permissive carbon sources. The unusual kinetic characteristics of YlPfk, and the intracellular concentrations of glycolytic intermediates during growth in glucose, suggest that YlPfk may play an important role in the regulation of glucose metabolism in Y. lipolytica, different from the role played by the enzyme in Saccharomyces cerevisiae.

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Assortment of carbon sources in medium for Yarrowia lipolytica lipase production: A statistical approach

Annals of Microbiology ◽

10.1007/s13213-014-0988-7 ◽

2014 ◽

Vol 65 (3) ◽

pp. 1495-1503 ◽

Cited By ~ 5

Author(s):

Agata Urszula Fabiszewska ◽

Danuta Kotyrba ◽

Dorota Nowak

Keyword(s):

Yarrowia Lipolytica ◽

Statistical Approach ◽

Carbon Sources ◽

Lipase Production

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A 37-amino acid loop in the Yarrowia lipolytica hexokinase impacts its activity and affinity and modulates gene expression

Scientific Reports ◽

10.1038/s41598-021-85837-8 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Piotr Hapeta ◽

Patrycja Szczepańska ◽

Cécile Neuvéglise ◽

Zbigniew Lazar

Keyword(s):

Amino Acid ◽

Yarrowia Lipolytica ◽

Carbon Sources ◽

Value Added ◽

Central Carbon Metabolism ◽

Cell Factory ◽

Loop Region ◽

Repressive Effect ◽

Depth Study ◽

Encoding Genes

AbstractThe oleaginous yeast Yarrowia lipolytica is a potent cell factory as it is able to use a wide variety of carbon sources to convert waste materials into value-added products. Nonetheless, there are still gaps in our understanding of its central carbon metabolism. Here we present an in-depth study of Y. lipolytica hexokinase (YlHxk1), a structurally unique protein. The greatest peculiarity of YlHxk1 is a 37-amino acid loop region, a structure not found in any other known hexokinases. By combining bioinformatic and experimental methods we showed that the loop in YlHxk1 is essential for activity of this protein and through that on growth of Y. lipolytica on glucose and fructose. We further proved that the loop in YlHxk1 hinders binding with trehalose 6-phosphate (T6P), a glycolysis inhibitor, as hexokinase with partial deletion of this region is 4.7-fold less sensitive to this molecule. We also found that YlHxk1 devoid of the loop causes strong repressive effect on lipase-encoding genes LIP2 and LIP8 and that the hexokinase overexpression in Y. lipolytica changes glycerol over glucose preference when cultivated in media containing both substrates.

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